Development of a new primer-probe energy transfer method for the differentiation of neuropathogenic and non-neuropathogenic strains of equine herpesvirus-1.

The research article presents the development of a new method to differentiate between neuropathogenic and non-neuropathogenic strains of the equine herpesvirus-1 through a novel approach known as the primer-probe energy transfer (PriProET) technique.

Background

• The equine herpesvirus-1 (EHV-1) is a major pathogen in horses causing anything from mild respiratory disease to severe neurological symptoms. There are different strains of this virus, some causing neurological symptoms (neuropathogenic) and others do not (non-neuropathogenic).
• Through comparative study, a single non-synonymous nucleotide change (A/G2254) in the open reading frame 30 (ORF30) region of the virus’s DNA was identified to be responsible for the change in the virus’s pathogenicity.
• Conventional methods for differentiating between these strains are not as efficient, leading to the necessity of a new technique.

Research Objective and Methodology

• The main aim of the study was to develop a new method of differentiation based on a technique known as primer-probe energy transfer (PriProET). This technique has previously been used for detecting and classifying various DNA and RNA viruses.
• This technique involves analysing the melting temperatures of these viruses. Variations in their genetic sequence will result in different peaks during the melting temperatures, thus enabling the researchers to differentiate between the different strains.

Key Findings

• The results strongly supported their hypothesis, with the melting temperature analysis showing exact correlations with the sequence variations of the targeted ORF30 region.
• This method easily identified the two genotypes (A/G2254) by the different peaks of their melting temperatures.

Conclusion

• The newly developed method was touted as simple, fast, specific and robust. It offered more flexibility compared to previous tests and showed potential to greatly aid in the identification and differentiation of various strains of the equine herpesvirus-1, and potentially other viruses.